This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have developed a microfluidic platform to generate stable gradients across 3D hydrogels and require confocal microscopy to image the cells as they migrate via chemotaxis toward EGF and SDF-1 gradients (FITC-dextran with a similar molecular weight). We will be using HT1080 (fibrosarcoma), MDA-MB-231 (metastatic breast cancer), and RAW (mouse macrophage) cell lines with GFP and RFP. In addition we will be conducting co-culture studies in our device in which the cells will be suspended in a 3D matrix (collagen, matrigel). We would like to also image the cell-cell interactions as well as the cell-matrix interactions (cell migration, degradation, proliferation, spreading, etc.) .